Bone formation by osteoblast-like cells in a three-dimensional cell culture.
Cells of the clonal osteogenic cell line MC3T3-E1 were seeded onto a three-dimensional matrix of denatured collagen type 1 and cultured for a period of up to 8 weeks. Specimens were analyzed by histological, enzyme histochemical, immunocytochemical, and ultrastructural methods and by in situ hybridization between day 7 and day 56 after seeding. In 56-day cultures, the MC3T3-E1 cells were arranged in a three-dimensional network and formation of bone-like tissue was indicated by calcification of a newly synthesized collagen type I matrix resembling osteoid and surrounding osteocyte-like cells. The differentiating culture showed high expression of osteocalcin and alkaline phosphatase activity. NIH3T3 fibroblasts used as control cells passed through the network of the substrate forming a confluent monolayer underneath. This culture system offers a potentially powerful model for bone formation in vitro and for investigating the osteogenic potential of bone-derived cells.
Casser-Bette
M
M
Abteilung für Molekulare Zellpathologie, Gesellschaft für Strahlen- und Umweltforschung (GSF), Neuherberg, Federal Republic of Germany.
Murray
A B
AB
Closs
E I
EI
Erfle
V
V
Schmidt
J
J
D000469:Alkaline Phosphatase / Q000378:metabolism; D001846:Bone Development; D002454:Cell Differentiation; D002460:Cell Line; D003094:Collagen / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D009693:Nucleic Acid Hybridization; D010006:Osteoblasts / Q000378:metabolism / Q000648:ultrastructure*
D003094:Collagen; D000469:Alkaline Phosphatase